Objective The aim of this experiment was to explore the effects of experimental intervention with normal saline on the microflora and short-chain fatty acid content in the cecum of broilers, and provide a theoretical basis for research on targeted regulation of the early intervention technique for broilers. Methods Eighty newly hatched chickens were randomly divided into two groups, namely the control group (group C) and the saline group (group S). The chickens in the group S were orally inoculated with 0. 5 mL of sterilized saline on the first two days after hatching, while the chickens in the group C were not treated with anything. Eight chickens from each group were randomly selected, weighed and their cecal contents were collected on days 3 and 7. Illumina Miseq high-throughput sequencing technology was used to determine the microfloral structure of the cecal content. The short-chain fatty acids contents in the ceca of broilers were determined by gas chromatography. Results Experimental intervention with normal saline showed no significant effect on the average daily weight gain of broilers in early developmental stages ( P > 0. 05). At the phylum level, the cecal flora profiles were similar between the two groups. Firmicutes, Proteobacteria and Bacteroidetes were the predominant phyla in the broiler ceca. At the genus level, compared with the group C, the relative abundance of Bacteroides and Cyanobacteria were increased by 160% and 143%, respectively, at the age of 3 days ( P < 0. 05). There was no significant difference in the relative abundance of cecal flora between the two groups at the age of 7 days. Furthermore, experimental intervention with normal saline dramatically reduced the acetic acid, butyric acid and isovaleric acid contents in the ceca of broilers aged 3 days ( P <0. 01). However, no significant difference was found at the age of 7 days ( P > 0. 05). Conclusions Experimental intervention with normal saline has an effect on the microflora and short-chain fatty acid contents in the ceca of broilers aged 3 days, but this effect is transient and may gradually disappear with increasing age.